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Coffee Break 7:30 AM - 8:00 AM
Scientific Symposium 100 8:00 AM - 10:00 AM Room: 201 C Cell and Gene Therapy in Cardiovascular Disease
Co-ChairsDavid A. Dichek, MD Darin J. Falk, PhD
Speakers
James M. Wilson, MD, PhD Gene Therapy for High Cholesterol Gene transfer to liver has the potential to treat a number of inherited dyslipidemias. Recent advances in the gene and protein regulatory pathways that control cholesterol metabolism suggest novel therapeutic targets which are amendable to therapeutic modulation via gene transfer. Progress in this area in animal models and potential applications in humans will be discussed.
Jay H. Traverse, MD Using a Network Approach to Conduct Cardiovascular Cell Therapy Trials – The CCTRN The Cardiovascular Cell Therapy Research Network (CCTRN) was established by the NHLBI to foster the development of cardiovascular cell therapy trials in the United States using a network approach. During its first 5 years, the CCTRN (Cleveland Clinic, University of Florida, Minneapolis Heart Institute, Texas Heart Institute, Vanderbilt University) and their satellites developed and completed 3 clinical trials using bone marrow mononuclear cells in the setting of acute myocardial infarction (TIME, LateTIME) and severe LV dysfunction (FOCUS-CHF). This talk will discuss the organization of the CCTRN, the benefits of using a Network approach to conduct clinical trials and the important development of core labs including a biorepository to study cell function.
H. Lee Sweeney, PhD Cardiac Gene Transfer for Inherited Cardiomyopathies Achieving efficient cardiac gene transfer in a large animal model has proven to be technically challenging. We are using percutaneous transendocardial injection catheter to deliver adeno-associated viral (AAV) vectors to the canine myocardium. With this injection route, AAV 6 is superior to the other serotypes evaluated in both dogs and rhesus macaques. We currently are trying to use the approach to treat the cardiomyopathy found Golden Retriver Muscular Dystrophy dog, the dystrophin-deficient dog model of Duchenne muscular dystrophy. The approach is to deliver AAV coding for a modified U7 small nuclear RNA (snRNA) carrying antisense sequence to target the exon splicing enhancers of exons 6 and 8 and correct the disrupted dystrophin reading frame. Delivery of this AAV-U7 vector results in expression cardiac dystrophin and improved cardiac function as assessed by cardiac magnetic resonance imaging (MRI).
Roger J. Hajjar, MD Novel Developments in Gene Therapy for Cardiovascular Diseases Chronic heart failure (HF) is a leading cause of hospitalization affecting nearly 6 million people in the U.S. with 670,000 new cases diagnosed every year. There is a desperate need for innovative therapies to reverse the course of ventricular dysfunction. One of the key abnormalities in both human and experimental HF is a defect in sarcoplasmic reticulum (SR) function, which controls Ca2+ handling in cardiac myocytes on a beat to beat basis. Over the last ten years Dr. Hajjar has undertaken a program of targeting important calcium cycling proteins in experimental models of heart by somatic gene transfer. This has led to the initiation and completion of phases 1 and 2 of the CUPID trial, which heralds a new era for gene therapy for cardiovascular disease.
Scientific Symposium 101 8:00 AM - 10:00 AM Room: 204 C Diverse Molecular Strategies from RNAi to Stem Cells to Correct Musculoskeletal Diseases
ChairJoel R. Chamberlain, PhD
Joel R. Chamberlain, PhD Virus-mediated RNAi Gene Silencing Strategies for FSHD and DM1 Therapy We are developing in vivo RNAi gene silencing approaches for dominant genetic disease of muscle. In a proof-of-principle study we demonstrated that AAV6-shRNAs prevent body-wide muscle weakness in a mouse model displaying some of the molecular and phenotypic features of facioscapulohumeral muscular dystrophy (FSHD). Our efforts are focused now on developing miRNA-based hairpin expression vectors targeting DUX4 mRNA, an mRNA closely linked to FSHD pathology. In a second application of viral RNAi-based gene silencing for treatment of muscular dystrophy we have improved human disease-related muscle electrophysiological defects and mRNA splicing alterations in the HSALR mouse model of myotonic dystrophy. Our findings highlight the potential for systemically delivered and sustained RNAi-based therapies for the dominant muscular dystrophies.
Frank Barry, PhD Stem Cell Therapy for Joint Repair Adult mesenchymal stem cells (MSCs) isolated from bone marrow and a variety of connective tissues have been extensively tested in the treatment of bone and cartilage repair and in osteoarthritis. In addition, fully allogeneic transplantation of human MSCs is tolerated in the immunocompetent host and allogeneic therapy has been effective in the treatment of graft-versus-host disease. However, there are many aspects of the biology of MSCs that are poorly described and a more exhaustive characterization is necessary to exploit these cells fully in the context of tissue repair. New approaches to understanding the biology of MSCs and the manner in which transplanted cells interact with the injured host will be described.
Darwin J. Prockop, MD, PhD Potential Therapies with Adult Stem/Progenitor Cells from Bone Marrow (MSCs) and the Proteins They Produce in Response to Signals from Injured Tissues We have recently used several different animal models of sterile inflammation to test the therapeutic benefits of MSCs and the proteins they produce in response to signals from injured tissues: myocardial infarction (Lee et al., PNAS 2009), zymosan-induced peritonitis (Choi et al., Blood 2011) and chemical injury to the eye (Oh et al., PNAS 2010). In each of these models MSCs reduced injury to the tissues by modulating excessive inflammatory responses that cause protease degradation of the tissues. The beneficial effects in each of the models were largely reproduced by administration of TSG-6, a multifunctional anti-inflammatory protein that is produced by MSCs and that modulates NF?B signaling in resident macrophages. Supported in part by NIH grants PO1 RR 17447, and 1R21EY020962.
Christopher H. Evans, PhD Gene Therapy for Arthritis This talk focuses on gene delivery to joints by intraarticular injection. This strategy promises to improve the treatment of numerous joint diseases, particularly rheumatoid arthritis and osteoarthritis. There is an abundance of supportive pre-clinical efficacy and safety data. A small number of clinical trials have taken place, and these will be described and discussed.
Scientific Symposium 102 8:00 AM - 10:00 AM Room: 203 AB Progress in Asia
Co-ChairsGuangping Gao, PhD Keiya Ozawa, MD, PhD
Shin-ichi Muramatsu, MD AADC Gene Therapy for Parkinson's Disease: Results of 3-5 Years of Follow-Up We started a phase I clinical study in 2007 to determine the safety of intrastriatal infusion of a recombinant AAV vector encoding human aromatic L-amino acid decarboxylase (AADC) in patients with Parkinson’s disease. At 36 months after treatment, all six patients showed improvements from baseline in off-medication Unified Parkinson’s Disease Rating Scale motor scores. One patient showed gradual cognitive decline over 48 months. One patient received bilateral deep brain stimulation of the subthalamic nucleus at 15 months after the procedure to reduce levodopa-induced dyskinesia that was alleviated but not abolished by gene therapy. Our findings provide class IV evidence showing the long-term safety and efficacy of AADC gene therapy for PD.
Zhijian Yang, MD, PhD Phase I & II Study of Adenovirus Vector Expressing HGF for the Treatment of Ischemic Heart Disease In Phase I Clinical Study, the 18 patients with ischemic heart disease were accomplished for adenovirus vector expressing HGF. The results showed that it was safe to use an adenovirus gene-transfer vector to deliver the human HGF gene by direct intracoronary injection. In Phase II Clinic Study, the 100 patients with ischemic heart disease will be selected, and 9 patients were treated with Ad-HGF by the catheter-based injection.
Yan Cheng, MD, PhD Phase I and Phase II Clinical Studies of an Oncolytic Adenovirus Expressing Granulocyte Macrophage Colony-Stimulating Factor for the Treatment of Head and Neck Cancers KH901 is a conditionally replicating oncolytic adenovirus that demonstrates selective intratumoral replication and necrosis in cancer patients. In the preclinical studies, KH901 showed strong tumor-cell selectivity in vitro and antitumor efficacy in the mouse models of human head/neck and hepatocellular cancer. In the phase I and phase II clinical studies, KH901 treatment was well tolerated for the cancer patients enrolled. While infectious KH901 was found in the urine samples, viral genomes and GM-CSF protein were detected in the serum samples. Most importantly, KH901 combined with chemotherapy prolonged the survival of the patients with late stage head and neck cancers. However, the strategy to improve the anti-tumor efficacy of KH901 (including better chemotherapy combination) need to be further developed.
Zhenhong Guo, PhD Phase II Study of Antigen-Stimulated Human DC as Therapeutic Vaccine Against Metastatic Colorectal Cancer Metastatic colorectal cancer is well known for its poor prognosis. This phase II clinical trial investigated the safety and efficacy of tumor antigen-pulsed dendritic cells vaccine combined with chemotherapy. One hundred and forty-two previously untreated patients with measurable metastatic colorectal cancer were randomly assigned to two treatment groups: tumor antigen-pulsed dendritic cells vaccine combined with chemotherapy treatment group and chemotherapy alone treatment group. The results showed that 45% patient in DC vaccine group, compared to 25% patients in control group, achieved partial response, indicating that antigen-pulsed DC vaccine is a promising therapeutic strategy for metastatic colorectal cancer.
Scientific Symposium 103 8:00 AM - 10:00 AM Room: 202 AB Progress and Prospects of Industry-sponsored Clinical Studies
ChairEstuardo Aguilar-Cordova, MDinf, PhD
Gabor Veres, PhD Gene Therapy Treatment for beta-Thalassemia: An Update Gene therapy for b-thalassemia has been challenging given the requirement for high level of hemoglobin production in a tightly controlled manner in hematopoietic cell lineage. We have previously published that following lentiviral b-globin gene transfer, an adult patient with severe ?E/?0-thalassaemia dependent on monthly transfusions has become transfusion independent. We will present and update on clinical efficacy and safety for up to 54 months on this first treated patient. In addition, we will provide initial safety and marking data on a second patient treated more recently with the same lentiviral vector. Preclinical safety, efficacy data using a modified Lentiglobin vector will be discussed together with future clinical plans to evaluate an improved version of the vector in ?-thalassaemia patients.
Philip D. Gregory, DPhil ZFN-edited CD4+ T cells for HIV/AIDS Therapy: Phase I Trials of SB-728-T in HIV-infected Subjects Zinc finger nucleases (ZFNs) provide a platform technology for the targeted engineering of the human genome. ZFNs targeting CCR5, an obligate co-receptor for entry of R5 HIV, render modified CD4 T cells resistant to HIV analogous to cells from subjects homozygous for a naturally occurring CCR5 mutation (CCR5?32). Here we report data from Phase I studies evaluating infusion of an autologous CCR5-modified T-cell product (SB-728-T) in HIV-infected subjects. We show that infusion was well tolerated, resulted in marked and prolonged enhancement of total circulating CD4 T-cell numbers, and prolonged engraftment of SB-728-T in peripheral blood and key tissue CD4 T-cell trafficking sites, such as the gut-associated lymphoid tissue. Importantly, following SB-728-T engraftment, a small cohort of treated HIV-infected subjects underwent experimental HAART interruption (TI), providing the first evidence of an impact on circulating HIV load and prompting confirmatory Phase 1/2 clinical trials.
Gabor M. Rubanyi, MD, PhD New Perspectives for Angiogenic Gene Therapy to Treat Myocardial Ischemia in Patients with Coronary Artery Disease Intracoronary administration of Ad5FGF4 (Generx) for the non-surgical treatment of myocardial ischemia in patients with recurrent angina pectoris has been studied in four previous clinical trials (AGENT). Several important insights for future directions have been drawn from these trials (representing the largest cardiovascular gene therapy clinical database to date) and also from recent animal research studies. One important discovery was the recognition of key role played by transient myocardial ischemia in augmenting both DNA-transfer efficiency and therapeutic efficacy. All of the new insights (clinical and experimental) have been incorporated in the clinical protocol of the ASPIRE pivotal phase 3 registration trial (now underway in the Russian Federation) and will be presented.
David J. Fink, MD Targeted Drug Delivery for Treatment of Intractable Pain Using Gene Transfer The use of analgesic drugs to treat severe pain is limited by off-target pharmacologic effects of these agents. We have created a series of nonreplicating herpes simplex virus-based vectors that, following injection into the skin, produce expression of analgesic peptides limited to a restricted set of peripheral sensory neurons. The results of the clinical trial of a nonreplicating HSV vector expressing preproenkephalin in patients with intractable pain from cancer will be presented.
Scientific Symposium 104 8:00 AM - 10:00 AM Room: 204 AB Solving, Evolving and Targeting Viral Capsids
ChairLuigi M. Naldini, MD, PhD
David V. Schaffer, PhD Molecular Engineering and Evolution of New Adeno-Associated Viruses for Gene Therapy Strong basic and translational efforts in the gene therapy field have culminated in successes in an increasing number of human trials and established the therapeutic potential of several viral vector systems. However, numerous challenges can limit the broader applicability of vectors such those based on adeno-associated virus (AAV) and lentivirus, including anti-vector humoral and cellular immunity, low transduction of some therapeutically relevant cells in vitro or in vivo, difficulty in overcoming cellular and physical barriers within complex tissue structures, and an inability for targeted delivery to specific cells. These challenges are not surprising, as nature did not evolve viruses for use as human therapeutics. We have been developing directed evolution approaches – which emulates the natural evolution process of iterative genetic diversification and selection – to create new “designer” AAV variants that address a number of these concerns. As recent examples, we have evolved AAV for the capacity to mediate high efficiency gene delivery to photoreceptors upon a simple injection into the vitreous humor of the eye, for the ability to evade human neutralizing antibodies, and for high efficiency gene delivery and gene targeting in adult and human pluripotent stem cells.
Aravind Asokan, PhD Towards a Synthetic AAV Vector Toolkit The adeno-associated virus (AAV) capsid is remarkably plastic. New strains have naturally evolved in different hosts driven by genetic drift and iterative mutagenesis. The overarching goal of our lab is to combine the tools and principles of molecular biology with chemistry & structural biology to generate a synthetic AAV vector toolkit. The resulting engineered viral strains are utilized to unravel viral infectious pathways and provide novel vectors for gene therapy.
Frederic D. Bushman, PhD DNA Integration in Human Gene Therapy The outcome of gene therapy can be tracked by deep sequencing of integrated vector genomes. This allows quantification of the contribution of different cell clones and molecular characterization of adverse events. The lecture will describe methods development and results from ongoing studies.
Florian Kreppel, PhD Understanding and Modulating (Adenovirus) Vector-Host Interactions by Combining Genetics and Chemistry The modification of the tropism viral gene transfer vectors has for a long time been based on genetic capsid alterations. Such modifications can allow for the introduction of novel receptor binding motifs and targeting. However, systemic vector delivery in vivo is still hampered by a multitude of different interactions of the vector capsids with various cellular and non-cellular host components, and this makes more extensive capsid modifications mandatory to achieve safe and efficient delivery. In this presentation using adenovirus (Ad) as an example, a capsid modification technology based on a combination of genetics and chemistry will be discussed. It will be shown how this technology allows to precisely modulate the liver tropism of Ad-based vectors including evasion from blood coagulation FX binding and Kupffer cell scavenging. Finally, a perspective on applying this technology for other non-enveloped vectors like AAV will be given.
Scientific Symposium 105 8:00 AM - 10:00 AM Room: 201 AB Update on CAR CD19 Studies
Jointly planned by the American Society for Blood and Marrow Transplantation
ChairMarcel van Den Brink, MD, PhD
Michel Sadelain, MD, PhD Targeting CD19 with CARs: Lessons Learned, So Far Chimeric antigen receptors effectively target T cells to tumors independently of HLA restriction. Based on the demonstration that human T cells targeted to CD19 could eradicate systemic, established B cell lymphomas and leukemias in immunodeficient mice (Brentjens, Nat Med, 2003), we initiated clinical trials in patients with chemorefractory chronic lymphocytic leukemia (CLL) and relapsed adult and pediatric acute lymphoblastic leukemia (ALL). Michel Sadelain will provide an update and comparison of results obtained to date.
Steven A. Rosenberg, MD, PhD Cancer Treatment Using Genetically Engineered T cells Cancer therapies have been developed based on the genetic modification of autologous T cells using genes encoding conventional alpha/beta T cell receptors (TCR) or chimeric receptors (CAR). Results of studies treating patients with solid cancers and lymphomas will be presented.
Carl H. June, MD CART19 Update for B cell Malignancy The recent clinical success of T cells redirected with chimeric antigen receptors (CARs) at several academic centers indicate this strategy as the new frontier for the immunotherapy of hematological malignancies. A number of issues facing the field will be addressed such as extending CAR T cell therapy beyond B cell malignancies, and whether CAR T cells may have a role in solid tumor therapy. Finally, approaches to control the fate of CAR T cells will be addressed.
Gianpietro Dotti, MD Update on Baylor College of Medicine CD19 CAR Studies Three phase I clinical studies using CD19-CAR T cells are current at Baylor College of Medicine. In the first, patients with B-cell derived malignancies are infused with two T-cell products that have been genetically modified with first generation or second generation (CD28 endodomain) CD19 CAR. This trial allows us to evaluate the specific role of CD28 costimulation in promoting CAR-T cell expansion and survival. Patients in the second study receive EBV-specific CTLs and activated T lymphocytes expressing a first generation and a second generation CD19 CAR, respectively. This trial allows us to evaluate the specific role of native TCR stimulation versus CD28 costimulation in promoting CD19 CAR-T cell expansion and survival. Finally, in the third clinical trial CD19 CAR modified CTLs are infused in patients receiving an allogeneic stem cell transplant. We generated monoculture CTL lines in which multivirus specific CTLs (CMV, EBV and AdV) are also rendered leukemia specific through the expression of CD19 CAR. This trial will evaluate whether these cells promote virus immune reconstitution and anti-leukemia activity without causing GvHD in recipients of allogeneic stem cell transplants.
Laurence JN Cooper, MD, PhD Sleeping Beauty System to Redirect Specificity of Clinical Grade T cells T cells can be genetically modified to prevent and treat malignancies. The Sleeping Beauty transposon and transposase system has been prepared for human application. This presentation will provide an update on the first-in-human application of this new approach to gene transfer to generate CD19-specific T cells.
Michael C. Jensen, MD Clinical Application of CAR Expressing T cells Derived from Purified Central Memory Precursors Despite considerable promise, adoptive therapy of B cell malignancies using CD19-CAR redirected T cells requires further refinement. A significant issue is the variability in the magnitude and duration of engraftment of transferred T cells, a key parameter that affects therapeutic efficacy. Based on insights related to the programming states of T cell subsets, it is clear that cell products derived from divergent T cell precursors are not uniform with respect to their subsequent engraftment fitness in the tumor bearing host. Our studies define an engraftment fitness hierarchy of CD8+ T cells (central memory>naïve>effector memory); accordingly, we have developed a GMP-compliant platform for immunomagnetic selection of central memory CD8 T cells and their subsequent activation, lentivirus transduction, and propagation to clinical trial cell doses. Our first-in-human central memory CD19CAR trial has been initiated in the setting of autologous HSCT for high risk B lineage lymphomas wherein T cells are infused shortly following myeloablation.
Break 10:00 AM - 10:30 AM
Education Session 110 10:30 AM - 12:00 PM Room: 202 AB Emerging Field Review: Advances in Tissue Engineering
In the past two decades, tissue engineering approaches have been relying on screening of biomaterials, biochemical and biophysical cues known to control cellular behavior. However, due to innumerous possible combinations, this trial-and-error approach has been non-efficient and limited successful. More recently, the notion that development and in vivo regeneration models can be used as a basis to design tissue engineering strategies has been increasingly appreciated. In this session, several examples will be presented on how in vitro and in vivo systems can be utilized to optimize tissue engineering strategies and how biomaterials and biophysical cues can be integrated to promote cell behavior towards tissue regeneration.
Co-ChairsJason A. Burdick, PhD Jeroen Eyckmans, PhD
Jason A. Burdick, PhD Tailoring Biomaterials to Modulate Cellular Interactions and Tissue Regeneration Biomaterials play an important role in tissue engineering, providing a 3-dimensional context to cellular positioning and tissue organization, as well as a range of biochemical and biophysical cues. Specifically, signals such as mechanical properties and the incorporation of adhesive ligands can regulate factors such as adhesion, proliferation, and differentiation of cells. More recently, these components are being explored in the context of dynamic presentation to mimic biological cascades and to modulate cellular behavior.
Robert L. Mauck, PhD Mechanical Stimulation to Direct Cellular Differentiation and Tissue Formation
Jeroen Eyckmans, PhD Utilizing In Vivo Models to Advance Musculoskeletal Tissue Engineering In the past two decades, tissue engineering approaches have been relying on screening of biomaterials, biochemical and biophysical cues known to promote stem cell differentiation. However, due to innumerous possible combinations, this trial-and-error approach has been non-efficient and limited successful. More recently, the notion that development and in vivo regeneration models can be used as a basis to design tissue engineering strategies has been increasingly appreciated. In this session, several examples will be presented on how insights gained from in vivo model systems can significantly advance the field of musculoskeletal tissue engineering.
Education Session 111 10:30 AM - 12:00 PM Room: 204 C Topical Review: iPS cells
Induced pluripotent stem cells (iPSCs) derived by reprogramming adult somatic cells is a new technology in the early phase of development that has promise for future cell therapies and opportunities for novel approaches to gene therapy for tissue and organ repair and for the treatment of inherited diseases. This session is designed to provide an update on this technology that includes discussion of methods of generating iPSCs and methods of genetic correction of inherited functional defects. The session has particular focus on differentiation of iPSC into hematopoitic cell lineages as a specific example of organ system studies made possible by iPSC technology.
ChairHarry L. Malech, MD
Linzhao Cheng, PhD Derivation and Gene Targeting of Human iPS Cells by Plasmid Vectors Dr. Cheng will give an overview of various methods for generating integration-free human iPS cells. Then he will emphasize on an improved method for generating integration-free iPS cells from blood mononuclear cells by transient expression of episomal vectors. He will discuss key issues of functional and genomic characterizations of human iPS cell lines. In addition, Dr. Cheng will discuss site-specific gene targeting or correction of mutations in human iPS cells using those from sickle cell disease as an example.
Cynthia Dunbar, MD Pluripotent Stem Cells and Genotoxicity Knowledge gained regarding the risks of genotoxicity conferred by integrating viral vectors and prolonged ex vivo culture from experience in the field of hematopoietic stem cell gene therapy informs concerns regarding induced pluripotent stem cells and other reprogrammed stem cell populations being developed for clinical applications. Many eventual applications of pluripotent cell populations for regenerative medicine will require genetic modification or correction of these cells, even if non-integrating approaches can be utilized for their generation. In this session, I will summarize the data to date regarding insertional mutagenesis and more general genotoxicity related to the generation and genetic modification of iPSCs, and current approaches being pursued to mitigate these risks.
Harry L. Malech, MD Use of iPS Cells for Studies of Chronic Granulomatous Disease We have determined an efficient method to derive iPS cell lines from the CD34+ hematopoietic stem cells present in 20 ml of peripheral blood. We have used this method to establish iPS cell lines from patients with both the X-linked and the most common autosomal recessive (47phox deficient) form of Chronic Granulomatous Disease. We have used zinc finger nuclease targeting to the AAVS1 site in the genome (adeno-associated virus 2 insert site) as a way to target a CGD corrective minigene into this “safe harbor” site. We show that this results in restoration of oxidase activity in neutrophils that we can differentiate from the gene-corrected CGD iPS cells. This presentation will discuss efficient generation of iPSC from small volumes of peripheral blood; the method to target minigene to the AAVS1 safe harbor site to functionally correct an inherited disorder (CGD); and the method we used to efficiently differentiate functional neutrophils from iPS cells.
Education Session 112 10:30 AM - 12:00 PM Room: 201 AB Topical Review: Lessons Learned from Cardiovascular Phase I/II Gene and Cell Therapy Trials
Gene and cell therapies have tremendous potential to provide highly specific, safe, and effective treatments for many different diseases, ranging from single gene defects to complex conditions that are primarily due to environmental causes. This session will present progress and prospects in cardiovascular gene and cell therapies and highlight some of the challenges that investigators have faced as they conduct gene and cell clinical trials.
ChairSonia I. Skarlatos, PhD
Jay H. Traverse, MD Ten Years of Cardiovascular Cell Therapy Trials – What have We Learned? Ten years ago German investigators described the safety and potential efficacy of administering autologous bone marrow-derived stem cells to patients following acute myocardial infarction (AMI). Over the next decade, multiple clinical trials have administered a variety of stem and progenitor cell products to patients with AMI, heart failure, chronic angina and critical limb ischemia. Although the safety of this approach has been well documented, the therapeutic efficacy of cell therapy has not been consistently demonstrated across the various trials and patient populations, in part due to patient selection, trial design and end-point analysis. This talk will focus on what we have learned in the last decade from cardiovascular stem cell trials and what is needed in future trials to advance the field.
H. Kirk Hammond, MD It's Harder Than You Think The strategy, issues, problems, and solutions of translating a basic research finding into a clinical gene transfer trial will be discussed. The presentation will embark with selection of a candidate gene and end with the day-to-day issues of sponsoring a multicenter clinical gene transfer trial. Discussion will include selection of appropriate preclinical models, conduct of biodistribution and toxicology studies, manufacture of clinical grade vector, design of a clinical trial, how to navigate the FDA approval process and write an IND application, and problems involved in the execution of such a clinical
Seppo Yla-Herttuala, MD, PhD, FESC Progress and Prospects of Cardiovascular Gene Therapy Trials for Ischemic Heart and Peripheral Vascular Diseases In this presentation recent developments in cardiovascular gene therapy will be presented. Also, vectors and current delivery strategies will be discussed. Main focus will be in angiogenic therapies.
Education Session 113 10:30 AM - 12:00 PM Room: 204 AB Topical Review: Regulatory RNAs
The role that non-coding RNA plays in regulating gene expression has become increasingly clear over the past decade. The first two talks in this session will discuss the mechanisms by which non-coding RNAs regulate gene expression at transcriptional and post-transcriptional levels. The final talk will discuss how regulatory RNAs are being developed for therapeutic purposes.
Co-ChairsLinda B. Couto, PhD Scott Q. Harper, PhD
Jodi L. McBride, PhD Post-transcriptional Gene Regulation: An Introduction to RNA Interference This session will introduce the basic principles of RNA interference (RNAi). We will discuss different species of small, silencing RNAs including short interfereing RNAs (siRNAs) short hairpin RNAs (shRNAs), micro RNAs (miRNAs) and piwi-interacting RNAs (piRNAs). A review of how small, silencing RNAs function endogenously to regulate gene expression will pave they way for following sessions in which we will discuss how the RNAi pathway can be exploited to silence genes of interest in both the laboratory and the clinic.
Kevin V. Morris, PhD The Emerging Paradigm of Long Non-Coding RNAs as Regulators of Gene Transcription and Epigenetic States Experimental observations have begun to highlight a role for long non-coding RNAs as modulators of epigenetic states in human cells. There are several proposed models, all of which appear to be highly informative and suggestive of an entirely new paradigm of gene regulation. These models and the implications of this body of work will be explored. Grant acknowledgments (NIAID R01AI084406 and NCI R01 CA151574).
Scott Q. Harper, PhD Therapeutic Applications of Regulatory RNAs For many years, the gene therapy field was primarily focused on developing gene replacement treatments for recessive disorders, while gene silencing strategies targeting dominantly inherited diseases lagged behind. This unequal focus largely arose from pragmatism: the molecular tools required for gene replacement strategies had been well-developed over the years, while those needed for silencing dominant disease genes were impractical or non-existent. Recent advancements in antisense RNA technology coupled with the discovery of RNA interference (RNAi) and its subsequent development as a gene silencing tool made it possible to begin developing efficient and targeted gene therapies for dominantly inherited diseases. This session will discuss potential therapeutic applications of RNA-based gene silencing strategies.
Education Session 114 10:30 AM - 12:00 PM Room: 201 C Topical Review: Synthetic Nucleic Acid Delivery Systems - From Bench to Bedside and Beyond
An exploration of some of the strengths, weaknesses, potential and limitations of synthetic systems for the delivery of nucleic acid-based drugs. An opening seminar on Strategic Considerations in the Development of Nucleic Acid-based Products from Paul Burke, a speaker who is uniquely positioned to give a talk describing what it takes to make a successful product, and why so many promising products have failed. This will be followed by two talks on synthetic delivery systems, how they can be best used in the laboratory and what to watch out for in designing and interpreting experiments utilizing these technologies.
ChairIan MacLachlan, PhD
Paul Burke, PhD Strategic Considerations in the Development of Nucleic Acid-Based Products Although the first synthetic antisense was approved in 1998 the oligonucleotide field has seen no subsequent approvals to date despite numerous clinical entries and countless research advances. Conventional drug development is an expensive, high-attrition endeavor; novel therapeutic modalities can entail additional risk and cost, especially when enabling technologies such as delivery systems are involved. In spite of this, the past two decades have witnessed a transformation of the pharmacopeia with the addition of complex biologics and products incorporating sophisticated delivery technologies. A historical retrospective of these successes provides lessons on strategic considerations relevant to the development of nucleic-acid based drugs.
Stephen L. Hart, PhD Lipid Mediated Delivery Systems Clinical trials with lipid-based delivery systems for gene and siRNA-mediated therapy of genetic diseases and cancers continue to make progress, and further opportunities are in advanced preclinical phases of development making this an exciting time for research in this area of gene therapy. Lipid-based vector systems encompass a great diversity of chemical structures and formulations that can provide alternatives to viral vectors in situations where their unique properties might be desirable such as their low immunogenicity profiles, or their packaging capabilities for large or small nucleic acids. Ultimately, however, efficiency of transfection is the key factor to wider clinical success of these delivery systems and advances are being made through research into areas such as improved lipid materials, methods of formulation into nanostructures with optimal biophysical properties and into ways of prolonging extracellular stability in vivo. Advances in our understanding of cellular transfection pathways from cell binding to nuclear or cytoplasmic deposition have led to the development of structures responsive to intracellular environmental triggers, such as pH changes in the endosomal compartment. Improvements in the DNA cargoes themselves such as CpG-free plasmids and minicircles are yielding improved levels of expression as well as more persistent expression, while developments in siRNA chemistries are advancing gene silencing efficiencies. Non-lipid components may also be combined with lipids to expand functionalities of nucleic acid formulations such as contrast agents for real-time imaging of biodistribution, targeting moieties for specificity, and stealth properties for prolonged persistence in vivo. Recent developments in these areas of research will be highlighted along with prospects for the future.
Ernst Wagner, PhD Polymeric Delivery Systems – Compacting, Shielding, Targeting and Intracellular Release In therapeutic siRNA or pDNA delivery, the different tasks at extra- and intra-cellular locations ask for dynamic carrier systems. Like natural viruses, such carriers have to be bioresponsive, sensing their environment and facilitating the next delivery step. Cationic polymers can complex and compact pDNA or siRNA into nanosized polyplexes. Functional modifications of polycations include PEG for shielding, ligands for receptor targeting, and endosome-destabilizing agents. Precise multifunctional pDNA and siRNA polyplexes can be designed by applying state-of–the-art synthetic chemistry, such as solid phase supported synthesis, and supramolecular assembly methods. E. Wagner, Polymers for siRNA delivery: Inspired by viruses to be targeted, dynamic and precise. Accounts Chem Res, online (2012). E. Wagner, Programmed drug delivery: Nanosystems for tumor targeting. Expert Opin. Biol. Ther. 7, 587-593 (2007).
Education Session 115 10:30 AM - 12:00 PM Room: 203 AB Topical Review: Recent Progress in Gene Therapy for CNS Disorders
The field of Gene Therapy has seen many recent developments concerning applications to treat diseases of the CNS. This session will deal with pre-clinical accomplishments concerning the transmission of vectors across the blood-brain barrier that should enable more widespread transduction of CNS targets via a less invasive delivery route and studies using novel vectors to treat CNS neurodegeneration. Recent significant progress in clinical trials for CNS neurodegenerative disease will also be presented.
ChairWilliam F. Goins, PhD
Matthew J. During, MD, PhD AAV-GAD Gene Transfer in Parkinson's Disease: A Progress Report A multi-center, randomized, double blind Phase 2 trial of AAV-GAD subthalamic gene transfer for advanced Parkinson's disease has been completed. The trial mets its 6 month primary endpoint, with data published in Lancet Neurology (vol. 10, pp309-319). Here, I review both the 6 month and open label 12 month data, include PET analysis. We show that the intervention is safe, without any serious adverse event, and is associated with highly significant improvements in both OFF UPDRSm scores as well as ON time at 12 months. Moreover, a treatment-related 2DG PET pattern correlates strongly with clinical outcome.
Guangping Gao, PhD AAV Transcytosis of the BBB The CNS is an important target for gene therapy of neurological disorders. Adeno-associated virus vector hold the promise for therapeutic gene transfer to treat a variety of diseases including the CNS disorders. For diseases that affect large areas of the CNS such as Canavan’s disease, administration of gene therapy vectors through vasculature to target the CNS globally is an attractive approach but hindered by the blood-brain-barrier (BBB). Recent progresses in developing novel AAV vectors to cross the BBB and target the CNS systemically as well as their preclinical applications will be discussed.
David S. Strayer, MD, PhD Strategies for Gene Delivery to the CNS to Treat HIV Encephalopathy Control of HIV-1 replication and related disease in the lymphoid organs and blood has greatly improved survival and quality of life, but most antiretroviral drugs penetrate the central nervous system (CNS) poorly, and so the CNS remains an important site of HIV-related tissue injury. Gene delivery strategies to protect the CNS from HIV-associated neurocognitive disorders (HAND) must take into account the facts that HIV-related neuronal loss occurs throughout the brain over an extended period of time and, particularly, the role of oxidant-related injury caused by HIV gene products in HIV-1 neurotoxicity. We devised strategies using recombinant SV40-derived gene transfer vectors to deliver antioxidant genes, in particular Cu/Zn superoxide dismutase (SOD1) and glutathione peroxidase (GPx1), to protect neurons throughout the CNS from the two principal neurotoxic HIV-1 proteins, namely envelope gp120 and Tat. Combining peripheral (intravenous) administration of mannitol with inoculation of SV(SOD1) and/or SV(GPx1) into the cerebrospinal fluid, we achieved transgene expression throughout the CNS at levels sufficient to provide safe, durable (=6 months) neuroprotection (=90%) from oxidant injury elicited by intracerebral inoculation of HIV-1 neurotoxins (gp120,Tat). Thus, delivering antioxidant transgenes throughout the CNS may provide an approach to preventing and treating long term HIV-related neurotoxic diseases, such as HAND.
Lunch Break (On Own - Not Provided) 12:00 PM - 1:15 PM
Education Session 120 1:15 PM - 2:45 PM Room: 203 AB Emerging Field Review: Cell and Gene Therapy for Skin Diseases
Epidermolysis bullosa is a family of skin adhesion disorders characterized by impairment of the dermal-epidermal junction, and characterized, in the non-lethal forms, by disfiguring blistering, recurrent infections, visual impairment, and an increased risk of cancer. Several therapeutic approaches have been proposed in the last decade, from autologous transplantation of cultured skin derived from genetically corrected epidermal stem cells to allogeneic fibroblast and bone marrow transplantation. Although preliminary proof of efficacy was obtained in most cases, feasibility and toxicity issues still limit full clinical development. Present hurdles and future hopes will be discussed for both cell and gene therapy approaches.
ChairFulvio Mavilio, PhD
Fulvio Mavilio, PhD Gene Therapy for Inherited Skin Adhesion Disorders Monogenic defects in the components of the dermal-epidermal junction lead to epidermolysis bullosa, a family of untreatable, severe skin adhesion diseases. Gene therapy based on transplantation of cultured skin derived from genetically corrected epidermal stem cells represents an attractive therapeutic option and a potential cure. Recent advances in gene transfer and stem cell culture technology are making this option more and more realistic. The progress and problems of gene therapy will be discussed in the context of the existing pre-clinical and clinical studies.
Jouni Uitto, MD, PhD Regenerative Medicine for Skin Disorders - The Paradigm of Heritable Blistering Diseases Heritable bullous diseases, as exemplified by epidermolysis bullosa (EB), manifest with blistering and erosions of the skin due to faulty adhesion of different cutaneous layers. Approaches of molecular therapy have been recently developed to counteract the clinical manifestations of these disorders. Ex vivo gene therapy, which utilizes transduction of patients’ keratinocytes in culture, with subsequent grafting back to the skin, has provided results attesting to the proof-of-principle. More recently, cell based therapy, consisting of injection of allogeneic fibroblasts into the blistering areas has shown promise for dermal repair. Finally, clinical trials of allogeneic bone marrow transplantation to the patients with EB have shown improvement in terms of lesser blistering and accelerated healing of the wounds. Refinement of these technologies are expected to lead to treatment of EB and other related skin fragility disorders in the near future.
Jakub Tolar, MD, PhD Stem Cell Transplantation for the Therapy of Skin Diseases One of the most fascinating and daunting challenges of current transplantation biology is to harness the potential of stem cells for tissue regeneration. Blood and marrow transplantation (BMT) has existed as the prototypical, anti-cancer stem cell therapy for more than 40 years, and can also be used for selected non-malignant lethal disorders, such as enzymopathies. Recently we have obtained laboratory and clinical evidence that BMT can also mediate tissue repair in congenital deficiency of skin extracellular matrix proteins, which results in a lethal blistering condition called epidermolysis bullosa (EB). Remarkably, healthy donor cells from the hematopoietic graft migrate to the injured skin. Simultaneously, there is an increase in the production of proteins that are deficient in EB, increased skin integrity, and a reduced tendency to blister formation. This new concept, whereby stem cells from one organ (bone marrow or cord blood) can heal another (skin and mucosa), can illuminate the mechanisms of cross-correction of a structural protein deficiency in extracellular matrix and regeneration of extramedullary tissues by hematogenous cells in other disease and injury states.
Education Session 121 1:15 PM - 2:45 PM Room: 202 AB Emerging Field Review: Humanized Animal Models
The topical review session, entitled “Humanized Animal Models ”, will focus on the development and utilization of humanized hematopoietic and hepatic murine model systems for basic research and preclinical trials for gene-therapy applications. The session will concentrate on the advantages and limitations of humanized mouse models as a means of studying the efficacy and safety of viral vectors in clinical trials. The session will be comprised of three talks.
ChairBruce E. Torbett, PhD, MSPH
Bruce E. Torbett, PhD, MSPH The Secret Life of Humanized Mice There is a basic and preclinical research need for small animal models that recapitulate human hematopoiesis and immune function. Immunodeficient mice “humanized” with human hematopoietic stem and progenitor cells, mature lymphocytes, human organs, and/or tissues, have proved beneficial for the study of cancer, gene delivery, hematopoiesis, infectious diseases, immunology, and transplantation. Advances in mouse strains that readily support human blood cell and tissue engraftment, the establishment and comparison of different humanized mouse models, and their uses, will be discussed.
J. Victor Garcia-Martinez, PhD Cell and Gene Therapy Applications of Humanized Mice Recent advances in the development of mice reconstituted with functional human hematolymphoid systems have made it possible to perform the in vivo evaluation of highly innovative cell and gene therapy experiments using human cells as targets. This presentation will cover the current state-of-the-art of these technologies. In addition, relevant aspects of the use of these models for the in vivo study of infectious diseases and cancer will be discussed.
Karl-Dimiter Bissig, MD, PhD Novel Human Liver Chimeric Mouse Models The mouse is a versatile small animal model, which has been well characterized and genetically modified towards our research needs. This lecture will focus on human liver chimerism and it’s most important applications in biomedical research. I will summarize the latest and most significant developments in the field of human liver chimerism and discuss the major challenges.
Education Session 122 1:15 PM - 2:45 PM Room: 204 AB Topical Review: Double Strand Break Repair and Targeted Genome Modification
Site-specific genome editing is a rapidly growing strategy in the gene therapy field. The key to this approach is to create site-specific damage to the genome which then activates the cell's endogenous repair machinery. In this session, pioneering work on the use of DNA double-strand breaks, single-strand DNA nicks, and rare cutting nucleases that formed the foundations for the genome editing field will be presented.
ChairMatthew H. Porteus, MD, PhD
David B. Roth, MD, PhD Nicks, Double-Strand Breaks and Gene Correction In this presentation I shall review recent work from several labs showing that homologous recombination and homology mediated gene targeting can be stimulated by site-specific nucleases that have been engineered to generate single-strand nicks. Such treatments may lower the background of nonhomologous, nontargeted integration events.
Maria Jasin, PhD Efficient Modification of the Genome by Double-strand Break Repair Chromosome breakage has serious consequences to the stability of the genome, hence, mammalian cells as others have multiple pathways with which to repair breaks to minimize the possibility of deleterious outcomes. Pathways of chromosome break repair can be co-opted, however, to induce genomic modifications, either by homologous recombination, to introduce a specific sequence modification, or non-homologous end-joining, to introduce a range of sequence modifications, including those that disrupt gene function. Chromosome translocations can also be engineered by this process. This presentation will discuss how chromosome break repair occurs and can be exploited for genome modification.
Andrew M. Scharenberg, MD Genome Editing with Rare Cleaving Nucleases Genome editing using rare cleaving nucleases is an emerging technology of potentially great significance to cell therapy. This presentation will provide an overview of the three major platforms of rare cleaving nucleases presently being applied therapeutically: homing endonucleases, zinc finger nucleases, and TAL effector nucleases. Topics to be covered include how to obtain or build nucleases of each class, approaches to vectorization of each class along with important caveats, and examples of therapeutic applications.
Education Session 123 1:15 PM - 2:45 PM Room: 201 C Topical Review: NIH Grant Writing and Career Development
This session will present NIH training and career opportunities for investigators at all different levels. In addition, a study section member and an applicant will discuss the do’s and don’ts of grant writing.
Terry R. Bishop, PhD Training and Career Development Opportunities from the NIH How can one gain support to pursue their research question? Dr. Bishop will lead you through a search of all funded NIH grants and show you how to find grants with similar topics as yours. This search may help you locate where in the NIH (in which Institute/Center of the NIH) your application will be most competitive. Secondly, Dr. Bishop will briefly discuss different types of grant activities, especially ones to be used during early career development stages, so you can find the grant mechanism most closely suited for your level of experience. After a brief discussion of the life cycle of a grant (process of applying for a grant), the medical/graduate school Loan Repayment Program will be described.
Brendan Lee, MD, PhD NIH Review: The Study Section Member’s Perspective Study Section review of scientific merit is an essential component of the NIH funding of gene and cell therapy studies. Factors that contribute to the evaluation of prospective applications will be reviewed and strategies for emphasizing the impact of proposed science will be discussed.
Katherine P. Ponder, MD The Do’s and Don’t of NIH Grant Writing This session will provide attendees with a practical approach to writing their first NIH grant. A critical starting point is to formulate the specific aims early, and to discuss them with as many people as possible to gather advice as to whether or not a particular aim is likely to be viewed favorably by the study section. The applicant needs to make sure that major points are made clear, and to convince the reader that he or she has the tools needed to go forward with the project. It is helpful to include some preliminary results to support the feasibility of the project, but do not go overboard, and make sure that all data that are presented are clearly described.
Education Session 124 1:15 PM - 2:45 PM Room: 204 C Topical Review: Vector Design for the Clinic
Vector Design for the Clinic. This session aims to provide answers to the following questions: How to optimize your vector of choice for optimal gene expression? What to take into consideration in order to maximize the safety? How can vector design impact regulatory approval? How to navigate the world of intellectual property when building a vector to the clinic? How does vector formulation and manufacturing concerns influence design?
Co-ChairsLuk H. Vandenberghe, PhD Hildegard Büning, PhD
Robin R. Ali, PhD AAV Vector Design for Ocular Gene Therapy Over the last 10 years AAV-based vectors have been used in many different proof-of-concept studies of ocular gene therapy using animal models for a variety of disorders from inherited retinal degeneration to uveitis. Recently AAV vectors have also been used in ocular gene therapy clinical trials. In this presentation I will review some of the issues surrounding the use of AAV vectors for effective gene therapy in the eye.
David B. Weiner, PhD Improved Plasmid Engineering and Delivery in Vivo The Delivery of Non Live, non replicating plasmids are an important tool for invivo gene delivery and immunization. However, this approach has suffered in its performance in larger animal models and in humans. Issues include that the genes delivered are only expressed at lower levels with in the cells of the body, as well as that transfection efficiencies, even with formulated plasmids, could be improved. We will discuss enhanced delivery technology as well as plasmid engineering and show examples of improved delivery and immunogenicity in response to these important technological advances.
Samuel C. Wadsworth, PhD
Break 2:45 PM - 3:15 PM
Oral Abstract Session 130 3:15 PM - 5:15 PM Room: 202 AB RNA Viral Vectors
Co-ChairsBoro Dropulic, PhD David W. Emery, PhD
Oral Abstract Session 131 3:15 PM - 5:15 PM Room: 201 C AAV Virus & Vector Biology
Co-ChairsMavis Agbandje-McKenna, PhD Luk H. Vandenberghe, PhD
Oral Abstract Session 132 3:15 PM - 5:15 PM Room: 204 C Chemical and Molecular Conjugates
Co-ChairsTheresa M. Reineke, PhD Millicent O. Sullivan, PhD
Oral Abstract Session 133 3:15 PM - 5:15 PM Room: 203 AB CNS: Metabolic
Co-ChairsJean Bennett, MD, PhD Douglas R. Martin, PhD
Oral Abstract Session 134 3:15 PM - 5:15 PM Room: 201 AB Musculo-skeletal Gene & Cell Therapy
Co-ChairsJeffrey Chamberlain, PhD Charles A. Gersbach, PhD
Oral Abstract Session 135 3:15 PM - 5:15 PM Room: 204 AB Stem Cell Therapy
Co-ChairsCarolyn Lutzko, PhD Eirini P. Papapetrou, MD, PhD
Exhibit Hall Open & Welcome Reception 5:15 PM - 7:15 PM Room: Hall A
New Member Welcome Reception 7:30 PM - 8:30 PM Room: Marriott 309-310
